Orientador: Iguatemy Lourenço Brunetti / Banca: João Olimpio Tognoli / Banca: Regina Célia Vendramini / Resumo: O uso de amostras-controle para acompanhamento do desempenho analítico é necessário para reproduzir a veracidade dos resultados das análises, pois monitora as variações que podem ocorrer no sistema analítico. O ideal para amostras-controle é ter a mesma matriz que as amostras dos pacientes, assim elas comportar-se-ão da mesma forma. Entretanto, para alcançar a estabilidade necessária, as amostras-controle passam por manipulações durante sua produção que podem alterar as propriedades da matriz. Dentre estas manipulações estão a adição de aditivos e mudanças físicas do meio como o congelamento ou liofilização. Neste estudo, nós produzimos amostras-controle com pool de soro humano trabalhados na forma líquida, congelados, conservados a ≤ -18°C e ≤ -80°C e sólida a 2 - 8°C liofilizados, sem preservante e com preservante e analisamos 23 analitos da rotina de laboratório clínico: ácido úrico, albumina, fosfatase alcalina, alanina aminotransferase, amilase, aspartato aminotransferase, bilirrubina direta, bilirrubina total, cálcio, capacidade latente de ligação do ferro, colesterol, creatinina, ferro, fósforo, gama-glutamil transferase, glicose, colesterol- HDL, desidrogenase lática, potássio, proteínas totais, sódio, triglicérides e uréia. As amostras foram analisadas por 300 dias, e comparado a estabilidade entre os soros sem preservante e com preservante nos diferentes processos de conservação. Os coeficientes de variação encontrados foram bastante satisfatórios, mas a estabilidade no decorrer do tempo foi melhor observada no soro com preservante, especialmente a 2 - 8°C - liofilizado, onde todos os analitos estudados permaneceram estáveis por 300 dias, com exceção do ácido úrico que foi de 240 dias / Abstract: Using control-samples for analytical performance monitoring is required to reproduce the veracity of the results of biochemical analyses, since them monitors changes that may occur in analytical system. The ideal for control-samples is to have the same matrix that samples of the patients, so they will behave the same way. However, to achieve the necessary stability control-samples undergo operations such as sum additives and/or physical changes of middle (freezing or lyophilization) during its production which can change the properties of the matrix. In this study, we produced control-samples with pool of human serum worked in frozen liquid form kept at ≤ -18°C and ≤ -80°C and freeze-dried solid kept between 2 - 8°C, without or with a preservative for 23 analytes of clinical laboratory routine: uric acid, serum albumin, alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, amylase, direct bilirubin, total bilirubin, calcium, latent iron-binding capacity, cholesterol, creatinine, iron, phosphorus, gamma glutamyl transferase, HDL-cholesterol, glucose, lactic dehydrogenase, total proteins, potassium, sodium, triglycerides and urea. The samples were analyzed for 300 days, and the stability compared between the without a preservative and with a preservative sera in different processes of conservation. The coefficient of variation found were quite satisfactory, but stability over time was better observed in with a preservative serum, mainly in serum stored between 2 - 8°C - freeze-dried, in which all analytes studied remained stable for 300 days, with the exception of uric acid that was for 240 days / Mestre

The field of clinical laboratory science is experiencing a critical shortage of qualified professionals. Because health care practitioners depend on the results of laboratory tests to help diagnosis and treat patients, it is important to address the current and future shortage in the laboratory workforce. There is limited research on factors affecting the turnover intentions of clinical laboratory employees. Accordingly, the research questions for this study examined the effect of burnout (BO) and organizational commitment (OC) on the turnover intention of laboratory employees in Florida. A cross-sectional survey design was used to examine the relationship between BO and OC on turnover intentions. Data were collected from licensed clinical laboratory directors, supervisors, technologists, and technicians using the following scales: demographic questionnaire, Maslach Burnout Inventory - General Survey, and Organizational Commitment Questionnaire. Linear regression and ANOVA were used to examine the relationships between these variables. The response rate was 18.4% (N = 184). Among clinical laboratory employees in Florida, the findings revealed significant predictive relationships between BO and turnover intention, OC and turnover intention, age and BO, and work shift and OC among clinical laboratory employees in Florida. Potential implications for positive social change from this study include reducing turnover among laboratory employees by allowing laboratory managers to create strategies that will reduce BO and increase OC, and thus decrease turnover intention.

The purpose of this study was to determine the impact of Regulation 1 of CLIA '88 on personnel needs in clinical laboratories of acute care hospitals in Virginia, resulting from proficiency testing, complexity level of testing and personnel standards. Because the legislation was enacted and passed with the intent of improving the quality of laboratory testing in every setting, the problem of the study was to determine the effects Regulation 1 of CLIA ’88 had on personnel needs for the delivery of quality clinical laboratory services in acute care hospitals of Virginia. A survey was sent to 140 acute care hospital laboratories in Virginia. There were 107 respondents, with 75 respondents providing usable data for this study. The remainder did not provide full laboratory services. Demographic information was obtained regarding bed capacity, educational levels of current personnel serving in different capacities, test volume, and percent of tests performed by different complexity levels. Comparisons were made among small, medium, and large facilities. The majority of respondents were representative of a facility with a bed capacity of 200 or less. The level of test complexity performed was similar regardless of the facility size. The majority of facilities did not anticipate an increase in personnel needs as a result of CLIA personnel standards. All facilities had personnel at all capacities that met required educational levels at this time. The majority of facilities did not anticipate an increase in personnel as a result of increased proficiency testing; however, when a projected need was indicated, there was a greater need indicated for AS degree level personnel followed by BS level personnel with a decline indicated in non degreed personnel. Staffing pattern changes related to increased proficiency testing indicated differences in the projected needs of small, medium, and large facilities. Barriers for implementing CLIA ’88 personnel standards were identified from the literature review and the pilot study. Respondents were asked to identify the barriers that were most Significant; they are in order as ranked: cost, availability of qualified personnel, and CLIA not reflecting the depth of knowledge and judgment needed to make independent competent judgment. The barriers were also reviewed by hospital bed size. It was concluded that as the number of tests being sent out increased, the number of tests being performed in-house have increased at the moderate complexity level, a level which requires less qualified personnel. The intent of the law to improve the quality of laboratory testing has not occurred in every setting. If the intent of the law is implemented, a need exists to provide educational opportunities at the AS and BS level for experienced personnel. Respondents did not perceive criteria as established by CLIA ‘88 as being adequate to determine the qualifications of personnel, who are responsible for quality patient test results in all settings. / Ed. D.

Retiring baby boomers and the lack of interest and awareness among college students to enroll in an accredited Clinical Laboratory Science (CLS) program have created a shortage of CLS professionals in the 21st century. The U.S. Bureau of Labor Statistics predicts 18,000 CLS vacancies by 2018. However, only about 5,000 students graduate from accredited CLS programs each year. The purpose of this study was to explore students' perceptions of allied health professions and factors that influenced students and CLS professionals to select CLS as a profession. Bandura's social cognitive career theory served as the theoretical framework for this phenomenological study. Convenient purposeful sampling was used to select the 7 CLS professionals, 5 high school students, and 5 college students in the Chicago area. Participants took part in either a 30- to 60-minute group session or a 45- to 90-minute semi structured interview. Qualitative analysis included open axial coding to identify emerging patterns and themes from the transcripts. Findings revealed that the perceptions of both high school and college students' knew little about the CLS profession, and factors influencing CLS as a career choice included interests in science, health care, and family. CLS professionals indicated their interests in science and a high demand for CLS services in the workforce led them to pursue careers in the field. Implications for social change include improving professional-development programs for student awareness of allied health professions and mitigating the shortage of clinical laboratory scientists.

Darbo tikslas. Palyginti darbo sąlygas privačiose ir viešųjų įstaigų klinikinėse laboratorijose. Tyrimo metodika. Tyrime dalyvavo 192 respondentai. 96 respondentai buvo vienos privačios X laboratorijos darbuotojai (penki padaliniai), kiti 96 viešųj�� įstaigų laboratorijų darbuotojai. Atsako dažnis 76,8 proc. Tyrimas atliktas 2007 m. spalio – 2008 m. sausio mėnesiais. Duomenys surinkti anoniminės anketinės apklausos būdu. Statistinė duomenų analizė atlikta naudojant kompiuterinį SPSS 13.0 statistinį paketą ir MS Excel. Rezultatai. Darbo aplinkoje su kenksmingomis cheminėmis medžiagomis, susiduria 97,9 proc. viešųjų ir 94,8 proc. privačios laboratorijos darbuotojai. Nepatenkinti darbo apsaugos priemonėmis buvo 55( 57,3 proc.) viešosios laboratorijos darbuotojų ir 25 (26 proc.) privačios laboratorijos darbuotojų. Geresniu užmokesčiu pasižymi privačių laboratorijų darbuotojai (p<0,05), o tai įtakoja ir tai, kad jie rečiau, nei viešųjų laboratorijų darbuotojai galvoja apie darbo keitimą. Didžiausias stresorius darbo aplinkoje buvo atsakomybė, už aliekamą darbą. (viešoje 82,3proc., privačioje 57,3 proc.) Statistiškai patikimiau (p<0,05) didesnę atsakomybę už atliekamą darbą jautė viešųjų laboratorijų darbuotojai. Labiausiai motyvaciją darbui didina atlygis, už padaryta darbą. Dažnesnį nuovargį darbo metu dažniau jaučia privataus sektoriaus darbuotojai, kurį įtakoja, didesni darbo krūviai, pamaininis darbas. Išvados. Privataus sektoriaus laboratorijų darbuotojai uždirba daugiau... [toliau žr. visą tekstą] / Final Paper Aim: Compare the work conditions in the public and private clinical laboratories. Work Methods: We have taken for the research work 192 respondents: 96 of them were employed in the private X laboratory (five departments) and the other 96 respondents were working in the public laboratories. Answer period 76,8 percent. This research was being run in the duration from the October, 2007 until the January, 2008. The questionnaire data have been gathered anonym. The statistic data analysis were put up by the help of statistical package SPSS and MS Excel. Results. In the environment with the toxic sanitary conditions work 97,9 percent of employees of public laboratories, and 94,8 percent of workers are engaged for the private laboratories. 55 (57,3 percent) of the public laboratories workers were not satisfied with the work protection conditions, and 25 (26 percent) of the private laboratory workers weren’t quite happy about the mentioned matter. A better income is assessable for the private laboratories employees (p<0,05), - this can admit them the more constant post of work than that of the public laboratories workers, who have to change their work therefore. The most ordinary factor of hectic in the environment was set the stressor of responsibility for the accomplished work as follows: 82,3 percent of the public sector respondents, and 57,3 percent of the private laboratories workers. The statistic reliable (p<0,05) responsibility for the work was shown by the... [to full text]

Neonatal alloimmune thrombocytopenia (NAIT) is caused by maternal alloantibodies against fetal platelet antigens inherited from the father and which are absent from maternal platelets. In Caucasians, antibodies against the Leu33 (HPA-1a) polymorphism of integrin β3 (part of the platelet αIIbβ3 complex) account for >70% of cases. Antenatal screening for these antibodies does not currently take place in the UK, partly because of the absence of sensitive, predictive tests. We hypothesized that the poor sensitivity and predictive abilities of current assays are due to the use of β3 in an inappropriate conformation, resulting in sub-optimal binding of HPA-1a antibodies. We hypothesized firstly that in vitro induced changes to αIIbβ3 might alter accessibility of the HPA-1a epitopes to alloantibodies, thus reducing assay sensitivity. Secondly, we hypothesized that HPA-1a antibodies are stimulated by, and preferentially recognise, β3 in association with αv, a molecule present on placental syncytiotrophoblasts, and that reactivity against platelet αIIbβ3 reflects only cross-reactivity with αvβ3. Our first hypothesis was proven by demonstrating that use of the cation chelating compound EDTA, used by many diagnostic laboratories as a component of assay reagents or present in blood samples as anticoagulant, resulted in significantly reduced assay sensitivity. These findings were confirmed in an international workshop. Support for our second hypothesis was provided by demonstrating enhanced reactivity of a small panel of examples of anti-HPA-1a against αvβ3 compared to αIIbβ3 and by molecular modelling data. We also showed that HPA-1a antibodies can inhibit platelet function by using a novel application of the ROTEM® delta thromboelastograph and an immunofluorescence assay in which we demonstrated blocking of platelet function using a monoclonal antibody, PAC-1, that binds only to activated αIIbβ3. These studies provide possible explanations for the poor sensitivity and predictive abilities of current assays and suggest further areas for research.

Haematological malignancies and their treatment lead to prolonged periods of severe thrombocytopenia (platelet count ≤ 50 x 10⁹/l). Despite the use of prophylactic platelet transfusions, haemorrhage remains an important complication during this thrombocytopenic period. Within a 30 day period up to 70% of patients have clinically significant haemorrhage (World Health Organization (WHO) grade 2 or above bleeding) and up to 10% have severe or life-threatening haemorrhage (WHO grade 3 or 4 bleeding). Hence our current management of these patients to prevent haemorrhage is sub-optimal. The aim of this thesis was to identify clinical and laboratory factors that may predict the risk of haemorrhage in patients with haematological malignancies and severe thrombocytopenia. This was achieved via several different study designs and assessed the effect of clinical and laboratory factors on any or clinically significant haemorrhage and their effect on intracranial haemorrhage. This thesis has demonstrated that there is no consensus on how bleeding is assessed and graded in this patient group. Also it showed that the absolute immature platelet number may be a better alternative to the total platelet count to guide administration of platelet transfusions. Female sex, a previous history of a fungal infection, a high C-reactive protein, a high white cell count, a low platelet count, anaemia, impaired renal function, and recent clinically significant haemorrhage were all found to be independent risk factors for haemorrhage. Patients who were in complete remission from their haematological malignancy had a much lower risk of bleeding.

As our understanding of cancer biology increases and novel therapies are developed, an increasing number of predictive biomarkers are becoming clinically available. Aberrant acetylation has been strongly linked to tumourigenesis and the modulation of acetylation through targeting histone deacetylase (HDAC) has led to the introduction of many HDAC inhibitors. To date, two have had regulatory approval for the treatment of cutaneous T cell lymphoma (CTCL). Modifications in chromatin control underpin the mechanism of action of HDAC inhibitors. A genome wide loss-of-function screen identified HR23B as a gene that governs sensitivity to HDAC inhibitors. HR23B shuttles ubiquitinated cargo proteins to the proteasome and elevated levels may contribute to cell death mediated by this pathway. It also governs cell sensitivity to drugs that act directly on the proteasome. HDAC inhibitors influence proteasome activity and there may be a synergistic interaction with proteasome inhibitors. HR23B and HDAC6 interact and HDAC6 may be a negative regulator of apoptosis and a positive regulator of autophagy and through its ability to down-regulate HR23B, may impact on the cellular outcome of HDAC inhibitor treatment. Expression of HR23B has been correlated with clinical response to HDAC inhibitors in a retrospective analysis of CTCL patients. The tissue expression of HR23B and the autophagy marker LC3 has been investigated and there may be a reciprocal relationship in their expression in some tumours which may provide prognostic information and patients with low HR23B expression but high levels of autophagy appear to have a particularly poor prognosis. Well designed, biomarker-driven prospective clinical trials are needed to clarify the predictive and prognostic roles of HR23B.

Stroke is a common and multifaceted disease that often involves motor deficits in the upper limb. This thesis investigated reliability and validity of existing clinical assessments of upper limb function in persons with post-stroke symptoms and in non-disabled controls. Study I was conducted in a clinical setting where the Motor Evaluation Scale of Upper Extremity in Stroke patients (MESUPES) was assessed in persons post-stroke by two physiotherapists selected from a group of four.  Study II-IV took place in a motion analysis laboratory with an optoelectronic system. Kinematic measures and clinical measures were used to investigate the validity of the Arm Posture Score (APS), the Finger-to-Nose test (FNT), and the Standardised Nine Hole Peg test (S-NHPT) in persons post-stroke and non-disabled controls.   The results showed that the MESUPES had a high inter-rater reliability while the concurrent validity was not fully confirmed. MESUPES has a maximum score of 58 and the minimal detectable change ranged from 5 to 8 for a confidence level of 80% and 95%. The Arm Posture Scores, which include either four or six arm movement variables, were able to distinguish between the affected and non-affected arms, as well as between the affected arm and the non-dominant arm of the controls. The total movement time of the FNT, which is a coordination test, was able to distinguish persons post-stroke from controls, at least at a group level. Movement smoothness, accuracy and compensation, obtained from kinematic analysis, were the most discriminative variables for the FNT. Smoothness was most strongly correlated with the timed FNT and had the greatest association with the variance of the timed FNT. For the S-NHPT, which is a dexterity test involving grasping and reaching, the movement times, smoothness and compensation discriminated between the stroke group and the control group. Persons post-stroke spent considerably more time in the grasp-related parts of the task compared to controls. Smoothness and upper limb impairments had the strongest correlation with the S-NHPT.   In conclusion, the clinical measures used within stroke rehabilitation seem valid and reliable, although some limitations are highlighted by the kinematic assessment. MESUPES was shown to be a reliable assessment of upper limb movement quality after stroke. The kinematic analysis revealed that the timed FNT does not have sufficient discriminative validity at an individual level. The timed FNT reflected speed-related aspects of pointing movements such as smoothness and length of the deceleration phase, but should not be used as an overall measure of upper limb coordination after stroke. The timed S-NHPT demonstrated sufficient discriminative validity and reflected smoothness and upper limb impairments. For both the FNT and S-NHPT, kinematic analysis showed that the clinical outcomes of those tests (time of performance) did not adequately detect qualitative aspects of the upper limb movements after stroke such as possible compensatory movements. Therefore, clinical assessments that capture qualitative aspects of upper limb movements would improve the assessment of upper limb coordination and dexterity after stroke. In addition, 3D movement analysis provided unique information about upper limb function after stroke, not least in persons with mild post-stroke impairments. The APS, for instance, which quantifies how much the arm swing during gait deviates from normal, discriminated between persons with stroke and non-disabled persons. Such subtle deviations are not possible to quantify with the human eye.

Angiogenesis, the growth of new blood vessels, is vital to tumour growth. Prevailing dogma has been that tumours cannot grow without angiogenesis. Based on this premise, anti-angiogenic drugs are used clinically. However, the principle of angiogenesis as an absolute requirement for tumour growth has been challenged with reports that many tumours are entirely or partially non-angiogenic. This study describes and quantifies characteristics of non-angiogenic non-small cell lung tumours, demonstrates non-angiogenic growth in small-cell/neuroendocrine lung tumours and investigates the underlying pathogenetic processes by comparison with angiogenic lung tumours. Hypoxia is an important stimulus for angiogenesis. Differences in response to hypoxia may determine whether a tumour produces new vessels. In order to test this, levels of. necrosis, often considered a surrogate marker of hypoxic stress, were quantified but no difference in quantity of necrosis was found Moreover, immunohistochemical investigation of hypoxia and angiogenesis factors provided no unambiguous explanation for the differences in angiogenesis. Significant differences were seen, however, in fibrosis and inflammation, which were both greater in angiogenic tumours. Differences were greater for lymphocytes rather than cells of the ‘innate’ immune system. This provided an alternative hypothesis: angiogenesis occurs during wound healing and in the growth of granulation tissue, so it is possible that tumour angiogenesis is a response to factors produced by immune cells rather than the tumour itself. A tumour’s angiogenic status may, therefore, be determined by the response it provokes from the immune system. Further work to test this theory would compare levels of immunogenic factors such as Tumour Necrosis Factor and tumour cell surface antigens such as the HLA class I molecules. The study concludes with an investigation into the molecular basis of non-angiogenic growth using the technique of comparative genomic hybridisation (CGH) which allows amplifications and deletions of areas of DNA to be calculated. High-resolution array CGH was evaluated against conventional CGH, and the results compared with previous RNA studies from our laboratory. These revealed a set of genes with consistent changes in both RNA and DNA, several of which form part of known angiogenic and inflammatory pathways.